Raydolfo M. Aprecio

The Antimicrobial Effect of MTAD: An In Vitro Investigation

Pulp and periradicular diseases are of microbial origin. To effectively clean the root canal system a disinfecting agent must be able to penetrate into difficult-to-reach areas and kill microorganisms with minimal damage to the host tissues. The purpose of this investigation was to test the ability of a mixture of a tetracycline isomer, an acid, and a detergent (MTAD) to kill Enterococcus faecalis and compare its efficacy to that of sodium hypochlorite (NaOCl) and ethylene diamine tetraacetic acid (EDTA). The zones of inhibition and minimum inhibitory concentrations were measured for these solutions. Measurement of zones of inhibition and determination of the minimum inhibitory concentrations showed that MTAD is as effective as 5.25% NaOCl and significantly more effective than EDTA (p < 0.0001). Furthermore, MTAD is significantly more effective in killing E. faecalis than NaOCl when the solutions are diluted (p < 0.0001). Measurement of the minimum inhibitory concentrations demonstrated that although MTAD is still effective in killing E. faecalis at 200x dilution, NaOCl ceases to exert its antibacterial activity beyond 32x dilution. EDTA did not exhibit any antibacterial activity. Based on the results of this study, it seems that MTAD is an effective solution in eradicating E. faecalis.

Minimum Contact Time and Concentration of Sodium Hypochlorite Required to Eliminate Enterococcus faecalis

INTRODUCTION The purpose of this investigation was to determine the concentration of sodium hypochlorite and the irrigation time required to disinfect dentin cylinders infected with Enterococcus faecalis. METHODS Four hundred fifty dentin cylinders (5 mm in diameter and 4 mm in height) with a lumen (2-3 mm in width) were prepared from freshly extracted bovine incisors. The cementum and predentin were then removed. The tubules were opened by using a 4-minute application with 17% ethylenediaminetetraacetic acid and 5.25% NaOCl and then exposed to E. faecalis (ATCC 4082) for 3 weeks in brain-heart infusion broth. The cylinders were then divided into 3 groups, and a 1.3%, 2.5%, or 5.25% concentration of NaOCl was applied in 5-, 10-, 15-, 20-, 25-, 30-, 35-, and 40-minute intervals for a total of 30 subgroups including positive and negative controls. Each test sample was placed into a tube of 2 mL brain-heart infusion broth and incubated for 72 hours. Absence of turbidity demonstrated no bacterial growth, whereas turbidity indicated presence of remaining viable bacteria. RESULTS The most effective irrigation regimen was 5.25% at 40 minutes, whereas irrigation with 1.3% and 2.5% NaOCl for this same time interval was ineffective in removing E. faecalis from infected dentin cylinders. CONCLUSIONS High concentration and long exposure to NaOCl are needed for elimination of E. faecalis contaminated dentin.

Biofilm removal by 6% sodium hypochlorite activated by different irrigation techniques

AIM To compare the removal of biofilm utilizing four irrigation techniques on a bovine root canal model. METHODOLOGY Fifty dentine specimens (2 × 2 mm) were infected with biofilm. The samples were then adapted to previously created cavities in the bovine model. The root canals were irrigated twice with 2 mL of 6% sodium hypochlorite for 2 min (4 min total). Following initial irrigation, the different treatment modalities were introduced for 60 s (3 × 20 s intervals). The evaluated techniques were needle irrigation, Endoactivator (Dentsply Tulsa Dental, Tulsa, OK, USA), passive ultrasonic irrigation and laser-activated irrigation (photon-induced photoacoustic streaming). The controls were irrigated with distilled water and conventional needle irrigation. Subsequently, the dentine samples were separated from the model and analysed using a scanning electron microscope (SEM). Fifteen operative fields were scanned per block, and SEM pictures were captured. Two calibrated evaluators examined the images and collected data using a four-degree scale. Nonparametric tests were used to evaluate for statistical significance amongst the groups. RESULTS The group undergoing laser-activated irrigation using photon-induced photoacoustic streaming exhibited the most favourable results in the removal of biofilm. Passive ultrasonic irrigation scores were significantly lower than both the Endoactivator and needle irrigation scores. Sonic and needle irrigation were not significantly different. The least favourable results were found in the control group. CONCLUSIONS Laser activation of 6% sodium hypochlorite significantly improved the cleaning of biofilm-infected dentine followed by passive ultrasonic irrigation.

Therapeutic Effect of TSG-6 Engineered iPSC-Derived MSCs on Experimental Periodontitis in Rats: A Pilot Study

Background We derived mesenchymal stem cells (MSCs) from rat induced pluripotent stem cells (iPSCs) and transduced them with tumor necrosis factor alpha-stimulated gene-6 (TSG-6), to test whether TSG-6 overexpression would boost the therapeutic effects of iPSC-derived MSCs in experimental periodontitis. Methods A total of 30 female Sprague-Dawley (SD) rats were randomly divided into four groups: healthy control group (Group-N, n = 5), untreated periodontitis group (Group-P, n = 5), iPS-MSCs-treated and iPSC-MSCs/TSG-6-treated periodontitis groups (Group-P1 and P2, n = 10 per group). Experimental periodontitis was established by ligature and infection with Porphyromonas gingivalis around the maxillae first molar bilaterally. MSC-like cells were generated from rat iPSCs, and transducted with TSG-6. iPSC-MSCs or iPSC-MSCs/TSG-6 were administrated to rats in Group-P1 or P2 intravenously and topically, once a week for three weeks. Blood samples were obtained one week post-injection for the analysis of serum pro-inflammatory cytokines. All animals were killed 3 months post-treatment; maxillae were then dissected for histological analysis, tartrate-resistant acid phosphatase (TRAP) staining, and morphological analysis of alveolar bone loss. Results Administration of iPSC-MSC/TSG-6 significantly decreased serum levels of IL-1β and TNF-α in the Group-P2 rats (65.78 pg/ml and 0.56 pg/ml) compared with those in Group-P (168.31 pg/ml and 1.15 pg/ml respectively) (p<0.05). Both alveolar bone loss and the number of TRAP-positive osteoclasts showed a significant decrease in rats that received iPSC-MSC/TSG-6 treatment compared to untreated rats in Group-P (p<0.05), Conclusions We demonstrated that overexpression of TSG-6 in rat iPSC-derived MSCs were capable of decreasing inflammation in experimental periodontitis and inhibiting alveolar bone resorption. This may potentially serve as an alternative stem-cell-based approach in the treatment and regeneration of periodontal tissues.

Effects of dietary fat and protein on DMH‐induced tumor development and immune responses

Although in three different mouse tumor systems with corn oil as dietary fat we previously found that milk protein decreased tumor development compared with beef, the results were reversed in 1,2-dimethylhydrazine (DMH)-injected mice. The purpose of this study was to determine if the latter result was due to the protein source. BALB/c mice (n = 280) were divided into five diet groups and injected 10 times at weekly intervals with DMH (20 mg/kg wt) or saline. Four diets contained 11% protein (casein, milk, or beef) and 5% fat (corn oil or beef tallow), and the AIN-76A diet was used as a control diet. The source of fat was a significant modulator of tumor development. Corn oil markedly increased total tumor volume and the number of tumors per mouse compared with beef tallow. Its tumor-enhancing effects were evident when it was combined with milk but not with casein. In addition, significantly lower lymphoproliferation and T-cell cytotoxicity against colon tumor cell targets was associated with corn oil consumption, whereas nonfat milk as the protein source was related to normal oxidative burst capacity of phagocytes. These results demonstrate that the source of dietary fat, in addition to the protein source, has a profound effect on both tumor development and immune responsiveness in this animal tumor system.

Comparison of topical ciprofloxacin to conventional antibiotic therapy in the treatment of experimental Pseudomonas aeruginosa keratitis.

We evaluated the efficacy of topical ciprofloxacin (3.0 mg/ml) in the treatment of experimental Pseudomonas aeruginosa keratitis in 60 rabbits. We compared ciprofloxacin treatment with double drug therapy consisting of tobramycin (13.6 mg/ml) plus polymyxin B (25,000 U/ml) or carbenicillin (6 mg/L). Two strains of P. aeruginosa were used. One was a strain well characterized for use in experimental Pseudomonas keratitis (ATCC organism 27853); the second was an organism from a patient with a Pseudomonas corneal ulcer. Rabbits were treated for 16 h, after which the corneas were excised, homogenized, and plated serially for residual colony-forming units. Both organisms responded significantly better to topical off-the-shelf ciprofloxacin than to therapy with two conventional antipseudomonal fortified antibiotic drugs (p =s 0.0001).

Modification of a transplantable colon tumor and immune responses in mice fed different sources of protein, fat and carbohydrate

The effects of different sources of dietary protein, fat and carbohydrate on tumor development and on tests relating to cell-mediated immunity were investigated in male BALB/c mice after subcutaneous injection of 8 X 10(4) 1,2-dimethylhydrazine (DMH)-induced colon tumor (no. 51) cells. Results indicated that mice fed the milk protein source (especially at the low protein level) had smaller tumors, a higher spleen cell proliferative response to stimulation by phytohemagglutinin (PHA), and greater cytotoxic T-cell activity against the tumor cells than those fed the comparable diets containing protein from the other sources. Peripheral blood lymphocytes only from the milk-fed mice, regardless of tumor presence, exhibited a relatively low response to PHA stimulation, thereby suggesting a dietary effect on the migration pattern of PHA-responsive lymphocytes. The level of protein significantly affected both T-cell and natural killer cell cytotoxicity. The tumor-bearing mice fed the diet containing sucrose (table sugar) had a significantly lower spleen cell response to PHA stimulation than those fed the comparable diet containing dextrin. The level or source of fat did not significantly affect any of the parameters tested in this system.

Lack of Virus Transmission by the Excimer Laser Plume

PURPOSE To test the possibility of pathogenic virus transmission into the operating suite during excimer laser treatment of corneal tissue. Such treatment vaporizes corneal tissue, which may put the surgeon at risk of infection from human immunodeficiency virus, hepatitis virus, or other viruses. We developed a model system to test the possibility of such virus transmission. METHODS Pseudorabies virus is a porcine enveloped herpesvirus similar in structure and life cycle to human immunodeficiency virus and herpes simplex virus. An excimer laser was used to ablate a virus-infected tissue culture plate while an uninfected tissue culture plate was in an inverted position over the infected plate. Six hundred laser pulses were applied. Pseudorabies virus in the excimer laser plume would, potentially, contact and infect the uninfected cells. The experiment was repeated 20 times with appropriate positive and negative controls. RESULTS None of the 20 uninfected plates was infected by the laser plume rising from ablation of infected tissue culture plates. Positive and negative controls performed as expected. CONCLUSIONS Even under conditions designed to maximize the likelihood of virus transmission, the excimer laser ablation plume does not appear capable of transmitting this particular live enveloped virus. Excimer laser ablation of the cornea of a human immunodeficiency virus (HIV)-infected or herpesvirus-infected patient is unlikely to pose a health hazard to the surgeon.

Efficacy of various disinfectants in killing a resistant strain of Pseudomonas aeruginosa by comparing zones of inhibition: implications for endoscopic equipment reprocessing

Objective:Previous studies have shown that high-level disinfection of GI endoscopes may not be reliably achieved using glutaraldehyde at room temperature. In our laboratory, we have isolated a strain of Pseudomonas aeruginosa that is resistant to disinfection with glutaraldehyde. We compared the bactericidal activity of various disinfectants against this organism.Methods:One hundred microliters of an overnight culture of this organism was spread onto blood agar plates. Twenty microliters of a disinfectant was placed on a sterile 7-mm filter paper, placed on the blood agar plate, and incubated overnight at 37°C to determine the zone of inhibition for each disinfectant tested. Disinfectants included Cidex, Dispatch, Virahol, OMNI II, Lysol, IodoFive, Lysol I.C. Spray, and Chlorox. The zone of inhibition (i.e., clearing) roughly correlates with the bactericidal strength of the disinfectant.Results:Compared with the glutaraldehyde-containing solution Cidex, the alcohol-containing disinfectants Lysol I.C. Spray and Virahol had the largest mean zones of inhibition (11.33 vs 20.60 and 20.55 mm; p= 0.0001). The hypochlorite compounds Chlorox (1:10 dilution) and Dispatch had mean zones of inhibition similar to that of Cidex (11.08 and 11.25 mm vs 11.33 mm; p= not significant). The phenolic compounds OMNI II and Lysol had mean zones of inhibition smaller than that of Cidex (10.50 and 10.35 mm vs 11.33 mm; p < 0.006), and the phosphoric acid and iodine–containing IodoFive had the smallest mean zone of inhibition (9.70 vs 11.33 mm; p= 0.0001).Conclusions:The alcohol-containing disinfectants had the largest zones of inhibition against resistant P. aeruginosa. These compounds may be more effective than glutaraldehyde for endoscopic equipment reprocessing.

Comparison of Efficacy of Pulverization and Sterile Paper Point Techniques for Sampling Root Canals

INTRODUCTION The purpose of this study was to compare the efficacy of the pulverization and sterile paper point techniques for sampling root canals using 5.25% NaOCl/17% EDTA and 1.3% NaOCl/MTAD (Dentsply, Tulsa, OK) as irrigation regimens. METHODS Single-canal extracted human teeth were decoronated and infected with Enterococcus faecalis. Roots were randomly assigned to 2 irrigation regimens: group A with 5.25% NaOCl/17% EDTA (n = 30) and group B with 1.3% NaOCl/MTAD (n = 30). After chemomechanical debridement, bacterial samplings were taken using sterile paper points and pulverized powder of the apical 5 mm root ends. RESULTS The sterile paper point technique did not show growth in any samples. The pulverization technique showed growth in 24 of the 60 samples. The Fisher exact test showed significant differences between sampling techniques (P < .001). The sterile paper point technique showed no difference between irrigation regimens. However, 17 of the 30 roots in group A and 7 of the 30 roots in group B resulted in growth as detected by pulverization technique. Data showed a significant difference between irrigation regimens (P = .03) in pulverization technique. CONCLUSIONS The pulverization technique was more efficacious in detecting viable bacteria. Furthermore, this technique showed that 1.3% NaOCl/MTAD regimen was more effective in disinfecting root canals.